Extracellular non-coding RNA signatures of the metacestode stage of Echinococcus multilocularis.
Extracellular RNAs (ex-RNAs) are secreted by cells through different means that may involve association with proteins, lipoproteins or extracellular vesicles (EV). In the context of parasitism, ex-RNAs represent new and exciting communication intermediaries with promising potential as novel biomarke...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2020-11-01
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| Series: | PLoS Neglected Tropical Diseases |
| Online Access: | https://doi.org/10.1371/journal.pntd.0008890 |
| _version_ | 1818728330104406016 |
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| author | María Eugenia Ancarola Gabriel Lichtenstein Johannes Herbig Nancy Holroyd Mara Mariconti Enrico Brunetti Matthew Berriman Krystyna Albrecht Antonio Marcilla Mara Cecilia Rosenzvit Laura Kamenetzky Klaus Brehm Marcela Cucher |
| author_facet | María Eugenia Ancarola Gabriel Lichtenstein Johannes Herbig Nancy Holroyd Mara Mariconti Enrico Brunetti Matthew Berriman Krystyna Albrecht Antonio Marcilla Mara Cecilia Rosenzvit Laura Kamenetzky Klaus Brehm Marcela Cucher |
| author_sort | María Eugenia Ancarola |
| collection | DOAJ |
| description | Extracellular RNAs (ex-RNAs) are secreted by cells through different means that may involve association with proteins, lipoproteins or extracellular vesicles (EV). In the context of parasitism, ex-RNAs represent new and exciting communication intermediaries with promising potential as novel biomarkers. In the last years, it was shown that helminth parasites secrete ex-RNAs, however, most work mainly focused on RNA secretion mediated by EV. Ex-RNA study is of special interest in those helminth infections that still lack biomarkers for early and/or follow-up diagnosis, such as echinococcosis, a neglected zoonotic disease caused by cestodes of the genus Echinococcus. In this work, we have characterised the ex-RNA profile secreted by in vitro grown metacestodes of Echinococcus multilocularis, the casuative agent of alveolar echinococcosis. We have used high throughput RNA-sequencing together with RT-qPCR to characterise the ex-RNA profile secreted towards the extra- and intra-parasite milieus in EV-enriched and EV-depleted fractions. We show that a polarized secretion of small RNAs takes place, with microRNAs mainly secreted to the extra-parasite milieu and rRNA- and tRNA-derived sequences mostly secreted to the intra-parasite milieu. In addition, we show by nanoparticle tracking analyses that viable metacestodes secrete EV mainly into the metacestode inner vesicular fluid (MVF); however, the number of nanoparticles in culture medium and MVF increases > 10-fold when metacestodes show signs of tegument impairment. Interestingly, we confirm the presence of host miRNAs in the intra-parasite milieu, implying their internalization and transport through the tegument towards the MVF. Finally, our assessment of the detection of Echinococcus miRNAs in patient samples by RT-qPCR yielded negative results suggesting the tested miRNAs may not be good biomarkers for this disease. A comprehensive study of the secretion mechanisms throughout the life cycle of these parasites will help to understand parasite interaction with the host and also, improve current diagnostic tools. |
| first_indexed | 2024-12-17T22:28:16Z |
| format | Article |
| id | doaj.art-34128e3bda1d4ebe9629a441bd91695b |
| institution | Directory Open Access Journal |
| issn | 1935-2727 1935-2735 |
| language | English |
| last_indexed | 2024-12-17T22:28:16Z |
| publishDate | 2020-11-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Neglected Tropical Diseases |
| spelling | doaj.art-34128e3bda1d4ebe9629a441bd91695b2022-12-21T21:30:17ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27271935-27352020-11-011411e000889010.1371/journal.pntd.0008890Extracellular non-coding RNA signatures of the metacestode stage of Echinococcus multilocularis.María Eugenia AncarolaGabriel LichtensteinJohannes HerbigNancy HolroydMara MaricontiEnrico BrunettiMatthew BerrimanKrystyna AlbrechtAntonio MarcillaMara Cecilia RosenzvitLaura KamenetzkyKlaus BrehmMarcela CucherExtracellular RNAs (ex-RNAs) are secreted by cells through different means that may involve association with proteins, lipoproteins or extracellular vesicles (EV). In the context of parasitism, ex-RNAs represent new and exciting communication intermediaries with promising potential as novel biomarkers. In the last years, it was shown that helminth parasites secrete ex-RNAs, however, most work mainly focused on RNA secretion mediated by EV. Ex-RNA study is of special interest in those helminth infections that still lack biomarkers for early and/or follow-up diagnosis, such as echinococcosis, a neglected zoonotic disease caused by cestodes of the genus Echinococcus. In this work, we have characterised the ex-RNA profile secreted by in vitro grown metacestodes of Echinococcus multilocularis, the casuative agent of alveolar echinococcosis. We have used high throughput RNA-sequencing together with RT-qPCR to characterise the ex-RNA profile secreted towards the extra- and intra-parasite milieus in EV-enriched and EV-depleted fractions. We show that a polarized secretion of small RNAs takes place, with microRNAs mainly secreted to the extra-parasite milieu and rRNA- and tRNA-derived sequences mostly secreted to the intra-parasite milieu. In addition, we show by nanoparticle tracking analyses that viable metacestodes secrete EV mainly into the metacestode inner vesicular fluid (MVF); however, the number of nanoparticles in culture medium and MVF increases > 10-fold when metacestodes show signs of tegument impairment. Interestingly, we confirm the presence of host miRNAs in the intra-parasite milieu, implying their internalization and transport through the tegument towards the MVF. Finally, our assessment of the detection of Echinococcus miRNAs in patient samples by RT-qPCR yielded negative results suggesting the tested miRNAs may not be good biomarkers for this disease. A comprehensive study of the secretion mechanisms throughout the life cycle of these parasites will help to understand parasite interaction with the host and also, improve current diagnostic tools.https://doi.org/10.1371/journal.pntd.0008890 |
| spellingShingle | María Eugenia Ancarola Gabriel Lichtenstein Johannes Herbig Nancy Holroyd Mara Mariconti Enrico Brunetti Matthew Berriman Krystyna Albrecht Antonio Marcilla Mara Cecilia Rosenzvit Laura Kamenetzky Klaus Brehm Marcela Cucher Extracellular non-coding RNA signatures of the metacestode stage of Echinococcus multilocularis. PLoS Neglected Tropical Diseases |
| title | Extracellular non-coding RNA signatures of the metacestode stage of Echinococcus multilocularis. |
| title_full | Extracellular non-coding RNA signatures of the metacestode stage of Echinococcus multilocularis. |
| title_fullStr | Extracellular non-coding RNA signatures of the metacestode stage of Echinococcus multilocularis. |
| title_full_unstemmed | Extracellular non-coding RNA signatures of the metacestode stage of Echinococcus multilocularis. |
| title_short | Extracellular non-coding RNA signatures of the metacestode stage of Echinococcus multilocularis. |
| title_sort | extracellular non coding rna signatures of the metacestode stage of echinococcus multilocularis |
| url | https://doi.org/10.1371/journal.pntd.0008890 |
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